Research Projects

The potential antitumor functions of Brassicaceae group vegetable crops, namely cabbage, broccoli, radish, Brussels sprouts and cauliflower are mainly due to the presence of isothiocyanates (ITCs) a derived products of glucosinolates (GSLs). Raphanus sativus (radish) possess large amounts of ITCs (Glucosinolates hydrolysis products) which serves functions as an antimicrobial, antimutagenic anticarcinogenic and potential antioxidant molecule (Nakamura et al. 2001). The major GSL includes glucoraphasatin (GRH; 4-methylthio-3-butenyl GSL; 80%) and glucoerucin (GER; 4-methylthiobutyl GSL; 10%) which are present in the seeds, roots, and shoots (Curtis 2011). Broccoli plants contain higher amounts of Glucoraphanin (GRA which hydrolysed and produce sulforaphane (SF))) due to 2-oxoglutarate dependant dioxygenase (AOP2) mutant gene which suggests the increased consumption of Broccoli vegetables provide protection against cancer and other diseases (Baskar et al., 2012 & 2016b). However, most of the Brassica crops possess least amount of GRA due to the presence of functional AOP2 gene. The high GRA was shown to present in a Japanese radish variety recently due to the occurrence of mutation in grs1 gene. Naturally derived bioactive compounds gains more attention due to the effective safer therapeutic values with least side effects and lower production costs as compared to synthetic drugs. Thus the proposed study is a composite approach includes, metabolic engineering of targeted knockout and transient methods for enhancing the production of potential chemopreventive GSLs (glucoerucin and glucoraphanin) in the edible crop R. sativus. Compared to control, SF treatment in mice suppressed the incedence and size of 4NQO-induced tongue tumors. NRF2-independent dephosphorylation/inactivation of pSTAT3A (oncogenic factor in HNSCC) was induced by the SF (Bauman et al., 2016). The outcome of the proposed project is expected to produce an elite radish cultivar with high SF using CRISPR-Cas based gene editing approach which could be useful to increase the revenue of the farmers and also to provide a healthier vegetable to the consumer. In addition elite hairy root transgenic (CRISPR-Cas KO grs1 mutants) will be selected and used for the larger production of SF at lab scale bioreactors. The extracted SF from in vitro produced plants will be examined for its potential against oral cancer under in vitro and in vivo studies. These valuable compounds would be used for the treatment as anti-cancerative and anti-inflammatory as well as antimicrobials. Industrial scale production of SF using this method will be economically feasible and sustainable format. Also the enhancement of SF in edible vegetable will enhance the intake of chemopreventive dietary molecule (SF) via their diet.